RNA silencing of integrin-linked kinase increases the sensitivity of the A549 lung cancer cell line to cisplatin and promotes its apoptosis.

Abstract

The expression of integrin-linked kinase (ILK) has been reported to be involved in the regulation of integrin-mediated processes, including cancer cell proliferation, migration and invasion. Previous studies have demonstrated that inhibition of ILK may be an underlying approach for treating cancer. However, whether the knock down of ILK affects growth and apoptosis of lung cancer cells remains to be elucidated. Importantly, whether downregulation of ILK increases the sensitivity of lung cancer cells to cisplatin and amplifies cell apoptosis also remains to be elucidated. In the present study, ILK downregulation was mediated by lentivirus-mediated RNA interference. The expression levels of associated genes were determined by reverse-transcription quantitative polymerase chain reaction and western blotting. Cell proliferation was evaluated using a modified 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and clone formation assay. The cell cycle and apoptosis were analyzed using flow cytometry. The current data revealed that lentivirus-mediated ILK gene silencing alone inhibited A549 cell proliferation and promotes cell cycle arrest, however, had no detectable effect on cell apoptosis. However, combined treatment with lentivirus-mediated ILK interference and cisplatin chemotherapy induced significantly more cell apoptosis than mono-chemotherapy or knockdown. The increased cell apoptosis and proliferation inhibition were attributed to abnormal downstream protein expression of ILK, including phospho-glycogen synthase kinase 3β, p-AKT, activator protein-1, β-catenin, cyclin D1 and matrix metalloproteinase-9. ILK inhibition may suppress the proliferation of A549 and increase A549 sensitivity to cisplatin. The combined treatment of ILK gene knockdown and chemotherapy has the potential to improve anticancer efficacy.

• Authors: Xiaozhen Zhao, Zhenye Xu, Zhongqi Wang, Zhonghua Wu, Yabin Gong, Lijuan Zhou, Yi Xiang
• Journal: Molecular medicine reports (Mol Med Rep) Vol. 12 Issue 1 Pg. 960-6 (Jul 2015) ISSN: 1791-3004 [Electronic] Greece
• PMID: 25760437 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antineoplastic Agents
• CCND1 protein, human
• CTNNB1 protein, human
• Transcription Factor AP-1
• beta Catenin
• Cyclin D1
• integrin-linked kinase
• GSK3B protein, human
• Glycogen Synthase Kinase 3 beta
• Protein Serine-Threonine Kinases
• Proto-Oncogene Proteins c-akt
• Glycogen Synthase Kinase 3
• Matrix Metalloproteinase 9
• Cisplatin

Topics

• Antineoplastic Agents (pharmacology)
• Apoptosis (drug effects)
• Cell Cycle Checkpoints (drug effects)
• Cell Line, Tumor
• Cell Proliferation (drug effects)
• Cisplatin (pharmacology)
• Cyclin D1 (genetics, metabolism)
• Drug Resistance, Neoplasm (drug effects, genetics)
• Epithelial Cells (drug effects, metabolism, pathology)
• Gene Expression Regulation, Neoplastic
• Genetic Vectors
• Glycogen Synthase Kinase 3 (genetics, metabolism)
• Glycogen Synthase Kinase 3 beta
• Humans
• Lentivirus (genetics, metabolism)
• Matrix Metalloproteinase 9 (genetics, metabolism)
• Protein Serine-Threonine Kinases (genetics, metabolism)
• Proto-Oncogene Proteins c-akt (genetics, metabolism)
• RNA Interference
• Respiratory Mucosa (drug effects, metabolism, pathology)
• Signal Transduction
• Transcription Factor AP-1 (genetics, metabolism)
• beta Catenin (genetics, metabolism)